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Time-resolved immunochromatography kit for synchronously detecting aflatoxin and carbaryl mixed pollution, and preparation method and application thereof

A time-resolved fluorescence and aflatoxin technology, applied in chemical instruments and methods, analytical materials, immunoglobulins, etc., can solve the problems of complex pretreatment steps and high cost of sample detection, and achieve simple methods, high sensitivity, and pre-sample The effect of simple processing

Active Publication Date: 2017-07-04
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have good stability, high sensitivity and good accuracy, but the pretreatment steps are complicated and the cost of sample detection is high

Method used

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  • Time-resolved immunochromatography kit for synchronously detecting aflatoxin and carbaryl mixed pollution, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 Obtaining of anti-carbaryl monoclonal antibody

[0039] Screening of hybridoma cell line Jnw1D2

[0040] 1. Animal immunization

[0041] Purchase 6 BALB / c mice aged 6 weeks, and use 6-(1-naphthyloxyformamide)-hexanoic acid (CNH) to couple bovine serum albumin (BSA) to obtain the complete antigen CNH-BSA of Carbaryl, Immunize mice. For the first immunization, complete carbaryl antigen was emulsified with an equal volume of Freund's complete adjuvant, and then injected subcutaneously at multiple points on the back of the neck of the mouse. The second immunization was carried out 3 weeks later, using Freund's incomplete adjuvant and equal volume of carbaryl complete antigen emulsification, and injected subcutaneously at multiple points on the back of the mouse. The third and fourth immunizations were separated from the last immunization by two weeks, and the immunization method was the same as that of the second time. The doses of the four immunizations we...

Embodiment 2

[0056] The acquisition of embodiment 2 anti-aflatoxin monoclonal antibodies

[0057] The anti-aflatoxin monoclonal antibody is produced by the secretion of the hybridoma cell line 1C11 with the preservation number CCTCC NO.C201018, specifically according to the method reported in the patent with the authorization number CN201010245095.5. The preparation method is: the obtained hybrid The tumor cell line 1C11 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, the ascites of the mice was collected, purified and treated to obtain anti-aflatoxin monoclonal antibodies. Among them, the purification method is the octanoic acid-ammonium sulfate method, and the specific operation is: filter the mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4°C, 12000r / min for more than 15min, absorb the supernatant, and dilute the supernatant with 4 times the volume Mix acetate buffer solution, slowly add n-octanoic acid while stirring...

Embodiment 3

[0059] Example 3 Simultaneous detection of aflatoxin, carbaryl time-resolved fluorescence kit and its application

[0060] Time-resolved fluorescence immunochromatography kit for the quantitative detection of aflatoxin and carbaryl, including fluorescent test strips, anti-aflatoxin monoclonal antibodies containing europium-labeled, europium-labeled carbaryl monoclonal antibodies, and sample reaction bottles , the fluorescent test strips such as figure 1 As shown, including the cardboard, one side of the cardboard is pasted with water-absorbing pads, detection pads and sample pads sequentially from top to bottom. Adjacent pads are overlapped and connected at the joints. The overlapping length is 1mm. Immunochromatography time-resolved fluorescent test strips The water-absorbing pad is 18 mm long and 4 mm wide; the detection pad is 25 mm long and 4 mm wide; the sample pad is 15 mm long and 4 mm wide, and the overlapping length of adjacent pads is 1 mm; the detection pad is based...

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Abstract

The invention relates to a time-resolved immunochromatography kit for synchronously detecting aflatoxin and carbaryl mixed pollution, and a preparation method and an application thereof. The kit comprises an immunochromatography time-resolved fluorescence test strip and a sample reaction bottle containing lyophilized products of an europium-labeled anti-aflatoxin monoclonal antibody and an europium-labeled anti-carbaryl monoclonal antibody, wherein the detection line of the immunochromatography time-resolved fluorescence test strip is respectively coated with an aflatoxin-bovine serum albumin conjugate and a carbaryl-ovalbumin conjugate; and the anti-carbaryl monoclonal antibody is secreted by a hybridoma cell strain Jnw1D2 with the preservation number of CCTCC NO.C201654. The kit can be used for synchronously detecting the content of aflatoxin and carbaryl in a sample, and has the characteristics of simplicity in operation, rapidness and high sensitivity.

Description

technical field [0001] The invention specifically relates to a time-resolved fluorescence immunochromatography kit for synchronous detection of mixed contamination of aflatoxin and carbaryl, a preparation method and application thereof. Background technique [0002] Aflatoxin is a major class of mycotoxin pollutants, with strong toxicity and wide pollution, and is classified as a class I carcinogen by the World Health Organization. Its toxicity is carcinogenic, mutagenic, and teratogenic to animals, and can cause DNA damage and immunosuppression. Aflatoxin pollution can occur in various links such as planting, harvesting, storage, transportation, and consumption. The contaminated products are mostly grains and nuts, and they are also detected in other products such as meat, eggs, and milk. Carbaryl is a class of carbamate pesticides widely used in vegetables, fruits, grains and other crops. It has a good control effect on pests. However, due to its abuse and prohibited use ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558
CPCG01N33/558G01N33/577G01N33/56961G01N2333/38G01N2430/10G01N2430/60G01N21/6408G01N21/8483G01N2021/7786C07K16/44C07K16/14G01N21/64G01N33/543G01N33/54388
Inventor 李培武唐晓倩张奇张兆威
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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